Chepyala Divyabharathi, Tsai I-Lin, Liao Hsiao-Wei, Chen Guan-Yuan, Chao Hsi-Chun, Kuo Ching-Hua
School of Pharmacy, College of Medicine, National Taiwan University, Taipei, Taiwan; The Metabolomics Core Laboratory, Center of Genomic Medicine, National Taiwan University, Taipei, Taiwan.
Department of Biochemistry and Molecular Cell Biology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan; Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan.
J Chromatogr A. 2017 Mar 31;1491:57-66. doi: 10.1016/j.chroma.2017.02.037. Epub 2017 Feb 21.
An increased rate of drug abuse is a major social problem worldwide. The dried blood spot (DBS) sampling technique offers many advantages over using urine or whole blood sampling techniques. This study developed a simple and efficient ultra-high-performance liquid chromatography-ion booster-quadrupole time-of-flight mass spectrometry (UHPLC-IB-QTOF-MS) method for the analysis of abused drugs and their metabolites using DBS. Fifty-seven compounds covering the most commonly abused drugs, including amphetamines, opioids, cocaine, benzodiazepines, barbiturates, and many other new and emerging abused drugs, were selected as the target analytes of this study. An 80% acetonitrile solvent with a 5-min extraction by Geno grinder was used for sample extraction. A Poroshell column was used to provide efficient separation, and under optimal conditions, the analytical times were 15 and 5min in positive and negative ionization modes, respectively. Ionization parameters of both electrospray ionization source and ion booster (IB) source containing an extra heated zone were optimized to achieve the best ionization efficiency of the investigated abused drugs. In spite of their structural diversity, most of the abused drugs showed an enhanced mass response with the high temperature ionization from an extra heated zone of IB source. Compared to electrospray ionization, the ion booster (IB) greatly improved the detection sensitivity for 86% of the analytes by 1.5-14-fold and allowed the developed method to detect trace amounts of compounds on the DBS cards. The validation results showed that the coefficients of variation of intra-day and inter-day precision in terms of the signal intensity were lower than 19.65%. The extraction recovery of all analytes was between 67.21 and 115.14%. The limits of detection of all analytes were between 0.2 and 35.7ngmL. The stability study indicated that 7% of compounds showed poor stability (below 50%) on the DBS cards after 6 months of storage at room temperature and -80°C. The reported method provides a new direction for abused drug screening using DBS.
药物滥用率上升是全球一个主要的社会问题。干血斑（DBS）采样技术相较于尿液或全血采样技术具有诸多优势。本研究开发了一种简单高效的超高效液相色谱 - 离子增强器 - 四极杆飞行时间质谱（UHPLC - IB - QTOF - MS）方法，用于分析使用DBS采集的滥用药物及其代谢物。本研究选取了57种涵盖最常见滥用药物的化合物作为目标分析物，这些药物包括苯丙胺类、阿片类、可卡因、苯二氮䓬类、巴比妥类以及许多其他新出现的滥用药物。采用80%乙腈溶剂，通过Geno研磨机进行5分钟萃取用于样品提取。使用Poroshell柱进行高效分离，在最佳条件下，正离子和负离子电离模式下的分析时间分别为15分钟和5分钟。对电喷雾电离源和包含额外加热区的离子增强器（IB）源的电离参数进行了优化，以实现所研究滥用药物的最佳电离效率。尽管这些滥用药物结构多样，但大多数滥用药物在IB源额外加热区的高温电离下表现出增强的质量响应。与电喷雾电离相比，离子增强器（IB）使86%的分析物检测灵敏度提高了1.5至14倍，并使所开发的方法能够检测DBS卡上的痕量化合物。验证结果表明，日内和日间精密度在信号强度方面的变异系数低于19.65%。所有分析物的萃取回收率在67.21%至115.14%之间。所有分析物的检测限在0.2至35.7 ng/mL之间。稳定性研究表明，在室温及 - 80°C下储存6个月后，7%的化合物在DBS卡上表现出较差的稳定性（低于50%）。所报道的方法为使用DBS进行滥用药物筛查提供了新方向。
